Actions of interferon tau during maternal recognition of pregnancy / Ações do interferon tau durante o reconhecimento materno da gestação

Maternal recognition of pregnancy in ruminants is a physiological process that requires an interaction between the conceptus and the mother in order to avoid luteal regression. Studies on 60s started to investigate the communication, and late on the 80s it was identified a Type I interferon called interferon tau. In the 90s the local action of interferon tau was described. It suppresses luteolytic pulses of prostaglandin F2 alpha inhibiting endometrial expression of estrogen and oxytocin receptors. Several studies presented the expression of interferonstimulated genes on extrauterine tissues. After, the endocrine action of interferon tau was described. This study revealed a greater interferon bioactivity in the serum of uterine vein from pregnant ewes when compared to nonpregnant ewes. Following a series of experiments were preformed to better understand the endocrine action of interferon tau in several extrauteine tissues. This review presents the endocrine action of interferon tau during maternal recognition of pregnancy period in ruminants.(AU)

O reconhecimento materno da gestação em ruminantes é um processo fisiológico que requer a interação entre o concepto e a mãe com o obejetivo de evitar a regressão luteal. Estudos na década de 1960 começaram a investigar essa comunicação entre concepto e mãe, e durante a década de 1980 foi identificado um tipo I de interferon que foi denominado interferon tau. Nos anos 1990, foi descrita a ação local do interferon tau. Essa ação suprime os pulsos luteolíticos de prostaglandina F2 alfa inibindo a expressão endometrial dos receptors de estrógeno e ocitocina. Muitos estudos têm apresentado a expressão dos genes estimulados por interferons nos tecidos extrauterinos. Posteriormente, a ação endócrina do interferon tau também foi descrita. Este estudo revelou uma maior bioatividade de interferon no soro da veia uterina de ovelhas prenhes quando comparada com ovelhas não gestantes. Após, uma série de experimentos foram realizados para melhor compreender a ação endócrina do interferon tau em vários tecidos extrauterinos. Esta revisão destaca a ação endócrina do interferon tau durante o período de reconhecimento materno da gestação em ruminantes.(AU)

Actions of interferon tau during maternal recognition of pregnancy / Ações do interferon tau durante o reconhecimento materno da gestação

Maternal recognition of pregnancy in ruminants is a physiological process that requires an interaction between the conceptus and the mother in order to avoid luteal regression. Studies on 60’s started to investigate the communication, and late on the 80’s it was identified a Type I interferon called interferon tau. In the 90’s the local action of interferon tau was described. It suppresses luteolytic pulses of prostaglandin F2 alpha inhibiting endometrial expression of estrogen and oxytocin receptors. Several studies presented the expression of interferonstimulated genes on extrauterine tissues. After, the endocrine action of interferon tau was described. This study revealed a greater interferon bioactivity in the serum of uterine vein from pregnant ewes when compared to nonpregnant ewes. Following a series of experiments were preformed to better understand the endocrine action of interferon tau in several extrauteine tissues. This review presents the endocrine action of interferon tau during maternal recognition of pregnancy period in ruminants.

O reconhecimento materno da gestação em ruminantes é um processo fisiológico que requer a interação entre o concepto e a mãe com o obejetivo de evitar a regressão luteal. Estudos na década de 1960 começaram a investigar essa comunicação entre concepto e mãe, e durante a década de 1980 foi identificado um tipo I de interferon que foi denominado interferon tau. Nos anos 1990, foi descrita a ação local do interferon tau. Essa ação suprime os pulsos luteolíticos de prostaglandina F2 alfa inibindo a expressão endometrial dos receptors de estrógeno e ocitocina. Muitos estudos têm apresentado a expressão dos genes estimulados por interferons nos tecidos extrauterinos. Posteriormente, a ação endócrina do interferon tau também foi descrita. Este estudo revelou uma maior bioatividade de interferon no soro da veia uterina de ovelhas prenhes quando comparada com ovelhas não gestantes. Após, uma série de experimentos foram realizados para melhor compreender a ação endócrina do interferon tau em vários tecidos extrauterinos. Esta revisão destaca a ação endócrina do interferon tau durante o período de reconhecimento materno da gestação em ruminantes.

Effects of ovarian pathologies and uterine inflammations on adenomyosis in bitches

Background: Adenomyosis is a non-neoplastic condition, characterized by the presence of endometrial glands or cells in the myometrium. Adenomyosis is defined by post-operative microscopic examination of uterine tissue and a rarely seen infertility problem in bitches. The process of transformation of healthy uterus into adenomiotic uterus is still a mystery. We aimed to determine role of ovarian pathologies and uterine inflammations on pathogenesis of uterine adenomyosis in bitches.Cases: After ovariohysterectomy, the uteri were fixed and processed by routine methods. Sections were stained with HematoxylinEosin, Massons trichrome and Periodic-Acid-Schiff and immunohistochemically; with p53 protein, Ki-67, α-smooth-muscle actin, cytokeratin-CK 8, 14, 19 antibodies, estrogen and progesterone receptors by Avidin-Biotin-Complex-Peroxidase method. Histopathologically, 7 of the collected tissue samples (n = 38) were diagnosed as adenomyosis. There were unilateral (n = 6) or bilateral (n = 1) adenomyosis in which endometrial glands had broken through the myometrium, and in some cases there were fibromyoma, inflammation, papillary hyperplasia and cysts in glands. Vaginal fibroma, fibromyoma and benign mix tumor (n = 1), simple carcinoma (n = 3), carcinosarcoma (n = 3) in mammary gland were determinated histopathologically. In addition to these findings, follicular cysts (n = 4), parovarian cyst (n = 1), adenocarcinoma and Rete ovarii adenoma were diagnosed in some ovaries. The CK 19 was scored at endometrial glands (n = 2) positively and also, muscle fibers were stained positively with α-smooth-muscle actin antibody (n = 7).Discussion: Adenomyosis is an infertility problem in adult dogs and is hard to detect with routine gynecological examinations. Adenomyosis can be seen unaccompanied by other gynecological problems such as endometritis or leiomyomas, while mammary tumors with adenomyosis have been previously reported in women and in only one case of a bitch.[...](AU)

Effects of ovarian pathologies and uterine inflammations on adenomyosis in bitches

Background: Adenomyosis is a non-neoplastic condition, characterized by the presence of endometrial glands or cells in the myometrium. Adenomyosis is defined by post-operative microscopic examination of uterine tissue and a rarely seen infertility problem in bitches. The process of transformation of healthy uterus into adenomiotic uterus is still a mystery. We aimed to determine role of ovarian pathologies and uterine inflammations on pathogenesis of uterine adenomyosis in bitches.Cases: After ovariohysterectomy, the uteri were fixed and processed by routine methods. Sections were stained with HematoxylinEosin, Masson’s trichrome and Periodic-Acid-Schiff and immunohistochemically; with p53 protein, Ki-67, α-smooth-muscle actin, cytokeratin-CK 8, 14, 19 antibodies, estrogen and progesterone receptors by Avidin-Biotin-Complex-Peroxidase method. Histopathologically, 7 of the collected tissue samples (n = 38) were diagnosed as adenomyosis. There were unilateral (n = 6) or bilateral (n = 1) adenomyosis in which endometrial glands had broken through the myometrium, and in some cases there were fibromyoma, inflammation, papillary hyperplasia and cysts in glands. Vaginal fibroma, fibromyoma and benign mix tumor (n = 1), simple carcinoma (n = 3), carcinosarcoma (n = 3) in mammary gland were determinated histopathologically. In addition to these findings, follicular cysts (n = 4), parovarian cyst (n = 1), adenocarcinoma and Rete ovarii adenoma were diagnosed in some ovaries. The CK 19 was scored at endometrial glands (n = 2) positively and also, muscle fibers were stained positively with α-smooth-muscle actin antibody (n = 7).Discussion: Adenomyosis is an infertility problem in adult dogs and is hard to detect with routine gynecological examinations. Adenomyosis can be seen unaccompanied by other gynecological problems such as endometritis or leiomyomas, while mammary tumors with adenomyosis have been previously reported in women and in only one case of a bitch.[...]

Partial ablation of endometrial glands in dogs after exposure to progestin during the neonatal period

Bitches with uteri devoid of endometrial glands should be sterile, and consequently could contribute to the population control of dogs. Considering that an inadequate exposure of the female reproductive system to steroids can lead to the formation of the uterine gland knock-out (UGKO) phenotype in some species, the aim of this study was to evaluate the effect of serial applications of medroxyprogesterone acetate (MPA) from birth until the age of six months on the development of endometrial glands in bitches. For this purpose, 16 female mongrel dogs from different litters were distributed into either an MPA group (n = 8), animals treated with 10 mg kg sc (Promone-E®, Pfizer, Brasil) at 3-week intervals, from day one after birth until the age of six months, or a control group (n = 8), composed of animals that only received a 0.9% NaCl solution in place of MPA. At six months of age, ovariohysterectomy was performed and uterine horn samples were collected for histological and immunohistochemical examinations. The bitches from the MPA-treated group presented a 35% decrease in the number of endometrial glands, a larger diameter of the endometrial glands, a greater epithelial height, as well as a greater thickness of the uterine wall, endometrium, and myometrium. However, no significant differences were observed between the two groups in the expression of ER-α, ER-β, and PR on the surface epithelium and endometrial stroma. Therefore, the serial application of MPA from birth until the age of 6 months do not completely ablate the development of the endometrial glands in bitches, but impair it by 35%.

Partial ablation of endometrial glands in dogs after exposure to progestin during the neonatal period

Bitches with uteri devoid of endometrial glands should be sterile, and consequently could contribute to the population control of dogs. Considering that an inadequate exposure of the female reproductive system to steroids can lead to the formation of the uterine gland knock-out (UGKO) phenotype in some species, the aim of this study was to evaluate the effect of serial applications of medroxyprogesterone acetate (MPA) from birth until the age of six months on the development of endometrial glands in bitches. For this purpose, 16 female mongrel dogs from different litters were distributed into either an MPA group (n = 8), animals treated with 10 mg kg sc (Promone-E®, Pfizer, Brasil) at 3-week intervals, from day one after birth until the age of six months, or a control group (n = 8), composed of animals that only received a 0.9% NaCl solution in place of MPA. At six months of age, ovariohysterectomy was performed and uterine horn samples were collected for histological and immunohistochemical examinations. The bitches from the MPA-treated group presented a 35% decrease in the number of endometrial glands, a larger diameter of the endometrial glands, a greater epithelial height, as well as a greater thickness of the uterine wall, endometrium, and myometrium. However, no significant differences were observed between the two groups in the expression of ER-α, ER-β, and PR on the surface epithelium and endometrial stroma. Therefore, the serial application of MPA from birth until the age of 6 months do not completely ablate the development of the endometrial glands in bitches, but impair it by 35%.(AU)

Endometrial transcription of microbial molecular patterns receptors in Gyr and F1 Holstein x Gyr postpartum cows / Transcrição endometrial de receptores de padrões moleculares microbianos em vacas Gir e F1 Holandês x Gir após o parto

Zebu and Holstein x Zebu crossbred have low incidence of uterine infection when compared to Holstein cows. Resistance to uterine infections may be associated with the ability to recognize invading microorganisms. Endometrial transcription of microbial molecular patterns receptors has been investigated in the postpartum period of Holstein cows, but it is completely unknown in Zebu or Holstein x Zebu cows. In this study, 9 Gyr and 12 F1 Holstein x Gyr cows were submitted to endometrial biopsies at the first and seventh days postpartum, with the objective to measure transcription levels of toll-like receptors (TLRs) 1/6, 2, 4, 5, and 9; nucleotide-binding oligomerization domain (NOD)-like receptors 1 and 2; and coreceptors cluster of differentiation 14 (CD14) and myeloid differentiation protein-2 (MD-2). There was a significant (P<0.05) decrease in transcription of TLR5 in Gyr, and an increase in transcription of TLR9 in F1 cows, between the first and seventh day postpartum. Both groups had low incidences of uterine infections up to 42 days postpartum. Uterine involution completed at 27.7 ± 10.1 and 25.1 ± 4.7 days postpartum for Gyr and F1 cows, respectively. In Gyr cows, higher transcription levels of TLR1/6 and NOD1 correlated to a longer period required for uterine involution. In F1 cows, lower levels of TLR1/6, TLR2 and NOD2 correlated to a longer period required for uterine involution. In conclusion, some pathogen recognition receptors associated significantly with the time required for uterine involution in Gyr and F1 cows.(AU)

Vacas Zebu e mestiças Holandês x Zebu apresentam baixas incidências de infecções uterinas quando comparadas às Holandesas. A resistência às infecções uterinas pode estar relacionada com a capacidade de reconhecimento dos microrganismos invasores. A transcrição endometrial de receptores de padrões moleculares microbianos tem sido investigada em vacas Holandesas recém-paridas, porém ainda é desconhecida em vacas Zebu e mestiças Holandês x Zebu. No presente estudo, nove vacas Gir e 12 F1 Holandês x Gir foram submetidas a biópsias endometriais no primeiro e no sétimo dia após o parto, com o objetivo de mensurar os níveis de transcrição gênica dos receptores tipo Toll (TLRs) 1/6, 2, 4, 5 e 9; receptores tipo NOD 1 e 2; e dos coreceptores CD14 e MD-2. Houve diminuição significativa (P < 0,05) do nível de transcrição de TLR5 em vacas Gir e aumento da transcrição de TLR9 em vacas F1, entre o primeiro e o sétimo dia após o parto. Os dois grupos apresentaram baixas incidências de infecções uterinas até 42 dias pós-parto. O período de involução uterina foi de 27,7 ± 10,1 e 25,1 ± 4,7 dias pós-parto, para vacas Gir e F1, respectivamente. No grupo de vacas Gir, altos níveis de transcrição de TLR1/6 e NOD1 tiveram correlação significativa com o prolongamento do período de involução uterina. No grupo de vacas F1, baixos níveis de transcrição de TLR1/6, TLR2 e NOD2 foram associados a maiores períodos de involução uterina. Portanto, os níveis de transcrição endometrial de alguns receptores de padrões moleculares microbianos na primeira semana após o parto podem estar relacionados com o tempo requerido para ocorrência da involução uterina em vacas Gir e F1.(AU)

Endometrial transcription of microbial molecular patterns receptors in Gyr and F1 Holstein x Gyr postpartum cows / Transcrição endometrial de receptores de padrões moleculares microbianos em vacas Gir e F1 Holandês x Gir após o parto

Zebu and Holstein x Zebu crossbred have low incidence of uterine infection when compared to Holstein cows. Resistance to uterine infections may be associated with the ability to recognize invading microorganisms. Endometrial transcription of microbial molecular patterns receptors has been investigated in the postpartum period of Holstein cows, but it is completely unknown in Zebu or Holstein x Zebu cows. In this study, 9 Gyr and 12 F1 Holstein x Gyr cows were submitted to endometrial biopsies at the first and seventh days postpartum, with the objective to measure transcription levels of toll-like receptors (TLRs) 1/6, 2, 4, 5, and 9; nucleotide-binding oligomerization domain (NOD)-like receptors 1 and 2; and coreceptors cluster of differentiation 14 (CD14) and myeloid differentiation protein-2 (MD-2). There was a significant (P<0.05) decrease in transcription of TLR5 in Gyr, and an increase in transcription of TLR9 in F1 cows, between the first and seventh day postpartum. Both groups had low incidences of uterine infections up to 42 days postpartum. Uterine involution completed at 27.7 ± 10.1 and 25.1 ± 4.7 days postpartum for Gyr and F1 cows, respectively. In Gyr cows, higher transcription levels of TLR1/6 and NOD1 correlated to a longer period required for uterine involution. In F1 cows, lower levels of TLR1/6, TLR2 and NOD2 correlated to a longer period required for uterine involution. In conclusion, some pathogen recognition receptors associated significantly with the time required for uterine involution in Gyr and F1 cows.(AU)

Vacas Zebu e mestiças Holandês x Zebu apresentam baixas incidências de infecções uterinas quando comparadas às Holandesas. A resistência às infecções uterinas pode estar relacionada com a capacidade de reconhecimento dos microrganismos invasores. A transcrição endometrial de receptores de padrões moleculares microbianos tem sido investigada em vacas Holandesas recém-paridas, porém ainda é desconhecida em vacas Zebu e mestiças Holandês x Zebu. No presente estudo, nove vacas Gir e 12 F1 Holandês x Gir foram submetidas a biópsias endometriais no primeiro e no sétimo dia após o parto, com o objetivo de mensurar os níveis de transcrição gênica dos receptores tipo Toll (TLRs) 1/6, 2, 4, 5 e 9; receptores tipo NOD 1 e 2; e dos coreceptores CD14 e MD-2. Houve diminuição significativa (P < 0,05) do nível de transcrição de TLR5 em vacas Gir e aumento da transcrição de TLR9 em vacas F1, entre o primeiro e o sétimo dia após o parto. Os dois grupos apresentaram baixas incidências de infecções uterinas até 42 dias pós-parto. O período de involução uterina foi de 27,7 ± 10,1 e 25,1 ± 4,7 dias pós-parto, para vacas Gir e F1, respectivamente. No grupo de vacas Gir, altos níveis de transcrição de TLR1/6 e NOD1 tiveram correlação significativa com o prolongamento do período de involução uterina. No grupo de vacas F1, baixos níveis de transcrição de TLR1/6, TLR2 e NOD2 foram associados a maiores períodos de involução uterina. Portanto, os níveis de transcrição endometrial de alguns receptores de padrões moleculares microbianos na primeira semana após o parto podem estar relacionados com o tempo requerido para ocorrência da involução uterina em vacas Gir e F1.(AU)

Cloprostenol administration in the first week postpartum reduces expression of oxytocin receptors in the endometrium in Holstein-Zebu cows / Administração de cloprostenol na primeira semana pós-parto pode reduzir a expressão de receptores de ocitocina no endométrio de vacas mestiças Holandês-Zebu

The present study investigated the hormonal profile and expression of prostaglandin F2α (PGF2α), oxytocin and estrogen receptors in uterine tissues of postpartum cows treated with cloprostenol. Twenty Holstein-Zebu crossbred cows were treated with saline solution (treatment CONT) or cloprostenol (treatment CLO), both administered two and five days postpartum. Blood samples were collected on days two, seven, 14, 21 and 28 postpartum for progesterone, PGF2α metabolite (PGFM) and estradiol determination, and endometrial biopsy was performed in order to quantify the expression of oxytocin receptor (OXTR), prostaglandin F receptor (PTGFR) and estrogen receptor 1 (ERS1) genes. In the CLO treatment, expression of OXTR was reduced (P<0.05) but no difference (P>0.05) between treatments was found for PTGFR and ERS1 expression. Estrogen concentrations increased progressively until day 14 (P<0.05) and the highest OXTR expression and lowest PTGFR expression were observed on day 14 (P<0.05) in both treatments. Serum PGFM concentrations were high throughout the experiment. In conclusion, cloprostenol administration at days two and five of postpartum seems to reduce OXTR expression in the endometrium in crossbred cows.(AU)

O presente estudo avaliou o perfil hormonal e a expressão gênica de receptores de prostaglandina F2α (PGF2α), ocitocina e estrógeno no endométrio de vacas pós-parto tratadas com cloprostenol. Vinte vacas mestiças Holandês-Zebu foram tratadas com solução salina (tratamento CONT, n = 10) ou cloprostenol (tratamento CLO, n = 10), ambos administrados dois e cinco dias após o parto. Amostras de sangue foram coletadas nos dias dois, sete, 14, 21 e 28 pós-parto para mensuração de progesterona, de metabólito de PGF2α (PGFM) e de estradiol, e foram obtidas biópsias endometriais para quantificar a expressão de PTGFR, OXTR e ESR1. No tratamento CLO, a expressão gênica de receptores de ocitocina foi menor (P<0,05). As concentrações de estrógeno aumentaram progressivamente até o dia 14 (P<0,05). A maior expressão de OXTR foi observada no dia 14 (P<0,05). A expressão de ESR1 foi semelhante entre os tratamentos (P>0,05). Os níveis de PGFM foram altos durante todo o estudo. Conclui-se que a administração de cloprostenol nos dias dois e cinco pós-parto parece diminuir a expressão de OXTR no endométrio em vacas mestiças.(AU)

Cloprostenol administration in the first week postpartum reduces expression of oxytocin receptors in the endometrium in Holstein-Zebu cows / Administração de cloprostenol na primeira semana pós-parto pode reduzir a expressão de receptores de ocitocina no endométrio de vacas mestiças Holandês-Zebu

The present study investigated the hormonal profile and expression of prostaglandin F2α (PGF2α), oxytocin and estrogen receptors in uterine tissues of postpartum cows treated with cloprostenol. Twenty Holstein-Zebu crossbred cows were treated with saline solution (treatment CONT) or cloprostenol (treatment CLO), both administered two and five days postpartum. Blood samples were collected on days two, seven, 14, 21 and 28 postpartum for progesterone, PGF2α metabolite (PGFM) and estradiol determination, and endometrial biopsy was performed in order to quantify the expression of oxytocin receptor (OXTR), prostaglandin F receptor (PTGFR) and estrogen receptor 1 (ERS1) genes. In the CLO treatment, expression of OXTR was reduced (P<0.05) but no difference (P>0.05) between treatments was found for PTGFR and ERS1 expression. Estrogen concentrations increased progressively until day 14 (P<0.05) and the highest OXTR expression and lowest PTGFR expression were observed on day 14 (P<0.05) in both treatments. Serum PGFM concentrations were high throughout the experiment. In conclusion, cloprostenol administration at days two and five of postpartum seems to reduce OXTR expression in the endometrium in crossbred cows.(AU)

O presente estudo avaliou o perfil hormonal e a expressão gênica de receptores de prostaglandina F2α (PGF2α), ocitocina e estrógeno no endométrio de vacas pós-parto tratadas com cloprostenol. Vinte vacas mestiças Holandês-Zebu foram tratadas com solução salina (tratamento CONT, n = 10) ou cloprostenol (tratamento CLO, n = 10), ambos administrados dois e cinco dias após o parto. Amostras de sangue foram coletadas nos dias dois, sete, 14, 21 e 28 pós-parto para mensuração de progesterona, de metabólito de PGF2α (PGFM) e de estradiol, e foram obtidas biópsias endometriais para quantificar a expressão de PTGFR, OXTR e ESR1. No tratamento CLO, a expressão gênica de receptores de ocitocina foi menor (P<0,05). As concentrações de estrógeno aumentaram progressivamente até o dia 14 (P<0,05). A maior expressão de OXTR foi observada no dia 14 (P<0,05). A expressão de ESR1 foi semelhante entre os tratamentos (P>0,05). Os níveis de PGFM foram altos durante todo o estudo. Conclui-se que a administração de cloprostenol nos dias dois e cinco pós-parto parece diminuir a expressão de OXTR no endométrio em vacas mestiças.(AU)

PAPEL DA HIPERPROLACTINEMIA E CLORIDRATO DE PIRIDOXINA NA EXPRESSÃO DE MEDIADORES HORMONAIS NO ÚTERO DE CADELAS COM PSEUDOCIESE

A pseudociese, também conhecida como pseudogestação é caracterizada como uma síndrome, em que cadelas não castradas apresentam sinais clássicos de peri e pós-parto. Os sinais clínicos vão desde alterações comportamentais, perda de apetite, perda de peso, vômito até o sinal mais evidente que é o aumento das glândulas mamárias e secreção de leite. A secreção láctea está relacionada com a redução dos níveis de progesterona plasmática associada ao aumento de prolactina. A prolactina exerce importante papel nas funções reprodutivas, no entanto, o excesso dessa substância está associado a alterações na fertilidade tanto em animais quanto em humanos, pela inibição da kisspeptina, peptídeo essencial para a reprodução, bloqueando o eixo hipotalâmico-hipofisário-gonadal (HPG). Para o tratamento farmacológico, as substâncias mais indicadas são os inibidores de prolactina, agonistas dopaminérgicos, a cabergolina e a bromocriptina. Uma alternativa terapêutica segura, capaz de reduzir os quadros de hiperprolactinemia em cadelas pseudogestantes é a vitamina B6 (cloridrato de piridoxina). Tanto os efeitos da hiperprolactinemia quanto do cloridrato de piridoxina sobre a expressão de mediadores hormonais no útero ainda é desconhecido. Acredita-se que a infertilidade causada pela hiperprolactinemia seja, além de origem central, no eixo HPG, como também de ação periférica, particularmente no útero. Nesse sentido, objetivou-se avaliar o efeito da hiperprolactinemia e do cloridrato de piridoxina na expressão de receptores de progesterona, estrógeno, andrógeno, hormônio tireoidiano, kisspeptina e KISS-1R, através da imunohistoquímica pela técnica da estreptavidina-biotina-peroxidase no útero de cadelas pseudogestantes. Foram utilizados fragmentos uterinos de 18 cadelas, divididos em três grupos: 1) controle não tratado, 2) tratado com cloridrato de piridoxina (50 mg/kg/dia; n=6) e 3) tratado com cabergolina (5µg/kg/dia; n=6). As cadelas com pseudogestação foram tratadas por 20 dias, submetidas a cirurgia de ovário-salpingo-histerectomia e os fragmentos uterinos foram fixados em formaldeído a 10% por 24 horas e depois processados pela técnica de inclusão em parafina. Avaliou-se a expressão de marcação nas regiões de epitélio endometrial, glândulas profundas e miométrio. A expressão de receptores de progesterona é maior em glândula profunda e miométrio nos grupos tratados (P <0,05). A expressão de receptores de estrógeno e KISS-1R é fraca nas regiões uterinas em todos grupos estudados (P > 0,05). Percebe-se expressão fraca ou nula de receptores de andrógeno em glândula profunda e miométrio nos grupos tratados (P < 0,001), resposta inversa a observada para progesterona. Nota-se expressão fraca de TR no grupo tratado com piridoxina em todas as regiões uterinas (P < 0,001) e expressão fraca de kisspeptina na superfície epitelial no grupo tratado com cabergolina

Pseudocyesis, also known as pseudogestation, is characterized as a syndrome, in which non-castrated bitches show classic signs of peri and postpartum. The signs range from behavioral changes, loss of appetite, weight loss, vomiting to the most obvious sign that the mammary glands are enlarged and milk is secreted. Milk secretion is related to the reduction in plasma progesterone levels associated with increased prolactin. Prolactin plays an important role in reproductive functions, however, the excess of this substance is associated with changes in fertility in both animals and humans, by inhibiting kisspeptin, a peptide essential for reproduction, blocking the hypothalamic-pituitary-gonadal axis (HPG ) For pharmacological treatment, the most suitable substances are prolactin inhibitors, dopaminergic agonists, cabergoline and bromocriptine. A safe therapeutic alternative capable of reducing hyperprolactinemia in pseudo-pregnant dogs is a vitamin B6 (pyridoxine hydrochloride). Both the effects of hyperprolactinemia and pyridoxine hydrochloride on the expression of hormonal mediators in the uterus are still unknown. It is believed that the infertility caused by hyperprolactinemia is not only central in the HPG axis, but also of peripheral action, particularly in the uterus. In this sense, the objective was to evaluate the effect of hyperprolactinemia and pyridoxine hydrochloride on the expression of progesterone, estrogen, androgen receptors, thyroid hormone, kisspeptin and KISS-1R, through immunohistochemistry by streptavidin-biotinperoxidase in the uterus in pseudostatic bitches. Uterine fragments of 18 dogs were used, divided into three groups: 1) untreated control, 2) treated with pyridoxine hydrochloride (50 mg / kg / day; n = 6) and 3) treated with cabergoline (5µg / kg / day ; n = 6). Pseudogestation bitches were treated for 20 days, submitted to ovariansalpingo-hysterectomy surgery and the uterine fragments were fixed in 10% formaldehyde for 24 hours and then processed using the paraffin inclusion technique. Marking expression is frequent in the regions of endometrial epithelium, deep glands and myometrium. The expression of progesterone receptors is greater in the deep gland and myometrium in the treated groups (P <0.05). An expression of estrogen receptors and KISS-1R is weak in the uterine regions in all groups studied (P> 0.05). The perception of weak or null expression of androgen receptors in the deep gland and myometrium in the treated groups (P <0.001), an inverse response to that observed for progesterone. There is a weak expression of TR in the group treated with pyridoxine in all uterine regions (P <0.001) and a weak expression of kisspeptin on the epithelial surface in the group treated with cabergoline.

Maintenance or regression of the corpus luteum during multiple decisive periods of bovine pregnancy

In ruminants, there are specific times during the estrous cycle or pregnancy when the corpus luteum (CL) may undergo regression. This review has attempted to summarize the physiological and cellular mechanisms involved in CL regression or maintenance during four distinct periods. The first period is near day 7 when animals that are ovulating after a period of low circulating progesterone (P4), such as first pubertal ovulation or first postpartum ovulation, are at risk of having a premature increase in Prostaglandin F2α (PGF) secreted from the uterus resulting in early CL regression and a short estrous cycle. The second period is when normal luteolysis occurs at day 18-25 of the cycle or when the CL is rescued by interferon-tau secreted by the elongating embryo. The uterine mechanisms that determine the timing of this luteolysis or the prevention of luteolysis have been generally defined. Induction and activation of endometrial E2 receptors result in induction of endometrial oxytocin receptors that can now be activated by normal pulses of oxytocin. Of particular importance is the observation that the primary mechanisms are only activated through local (ipsilateral) and not a systemic route due to transfer of PGF from the uterine vein to the ovarian artery. In addition at the CL level, studies are providing definition to the cellular and molecular mechanisms that are activated in response to uterine PGF pulses or pregnancy

Maintenance or regression of the corpus luteum during multiple decisive periods of bovine pregnancy

In ruminants, there are specific times during the estrous cycle or pregnancy when the corpus luteum (CL) may undergo regression. This review has attempted to summarize the physiological and cellular mechanisms involved in CL regression or maintenance during four distinct periods. The first period is near day 7 when animals that are ovulating after a period of low circulating progesterone (P4), such as first pubertal ovulation or first postpartum ovulation, are at risk of having a premature increase in Prostaglandin F2α (PGF) secreted from the uterus resulting in early CL regression and a short estrous cycle. The second period is when normal luteolysis occurs at day 18-25 of the cycle or when the CL is rescued by interferon-tau secreted by the elongating embryo. The uterine mechanisms that determine the timing of this luteolysis or the prevention of luteolysis have been generally defined. Induction and activation of endometrial E2 receptors result in induction of endometrial oxytocin receptors that can now be activated by normal pulses of oxytocin. Of particular importance is the observation that the primary mechanisms are only activated through local (ipsilateral) and not a systemic route due to transfer of PGF from the uterine vein to the ovarian artery. In addition at the CL level, studies are providing definition to the cellular and molecular mechanisms that are activated in response to uterine PGF pulses or pregnancy…(AU)

EFEITO DA RETENÇÃO DE PLACENTA E MASTITE CLÍNICA PÓS-PARTO NA EFICIÊNCIA REPRODUTIVA E EXPRESSÃO GÊNICA DE CITOCINAS E RECEPTORES ESTEROIDOGÊNICOS NO ENDOMÉTRIO DE VACAS LEITEIRAS MESTIÇAS

As doenças uterinas e não-uterinas no pós-parto impactam na eficiência reprodutiva 10 de vacas leiteiras. No entanto, além de pouco conhecimento em relação a vacas 11 mestiças, ainda não está claro como estas enfermidades interferem negativamente 12 na fertilidade. A hipótese deste estudo foi que vacas mestiças com retenção de 13 placenta (RP) e mastite clínica no pós-parto apresentam piores índices reprodutivos 14 por estas patologias interferirem na imunidade inata endometrial e expressão de 15 receptores dos hormônios esteroidogênicos ao final do puerpério. Portanto, os 16 objetivos com este estudo foram: a) avaliar o efeito da RP e mastite clínica pós17 parto na eficiência reprodutiva, período de serviço e intervalo parto-primeira 18 inseminação artificial (IA) de vacas mestiças leiteiras (experimento 1); b) avaliar o 19 efeito destas doenças na imunidade inata endometrial e receptores dos hormônios 20 esteroidogênicos (experimento 2). Para o exp. 1, 232 vacas foram avaliadas e 21 divididas nos grupos saudável, RP e mastite clínica. As vacas foram inseminadas à 22 medida que se observava estro ou submetidas ao protocolo de IATF. As avaliações 23 de ocorrência das doenças se davam após o parto até o final do período voluntário 24 de espera e o diagnóstico de gestação em torno de 32 dias após IA. Para o exp. 2, 25 foram utilizadas 30 vacas, divididas em três grupos (n=10) iguais ao experimento 1. 26 Entre 40-50 dias pós-parto era realizado exame clínico, exame ginecológico e 27 coleta de amostra endometrial por cytobrush para citologia endometrial e expressão 28 gênica pela técnica de qRT-PCR de IL1, IL6, CCL5, ESR1 e PGR. No exp. 1, a 29 taxa de concepção na primeira IA e porcentagem de vacas gestantes aos 150 dias 30 foram menores em animais doentes (RP, mastite) e os dias do intervalo parto31 primeira IA e período de serviço foram maiores nos animais doentes. No exp. 2, 32 26,7% (8/30) dos animais apresentaram endometrite subclínica. A IL1 apresentou 33 maior expressão nos grupos doentes, IL6 apresentou menor expressão no grupo RP 36 34 e a CCL5 não teve diferença entre os grupos. Não foi observado diferença na 35 expressão gênica dos receptores de esteroides ESR1 e PGR entre vacas doentes e 36 saudáveis. Concluiu-se que vacas com retenção de placenta e mastite clínica no 37 pós-parto apresentam piores índices reprodutivos, alteração na expressão gênica 38 endometrial das citocinas IL1 e IL6, porém não apresentam alteração na expressão 39 de receptores dos hormônios esteroidogênicos ao final do puerpério.

The postpartum uterine and non-uterine diseases impact the reproductive efficiency 44 of dairy cows. However, in addition to little knowledge regarding crossbred cows, 45 it is not yet clear how these diseases negatively affect fertility. The hypothesis of 46 this study was that crossbred cows with placenta retention (RP) and clinical mastitis 47 in the postpartum period have worse reproductive rates because these pathologies 48 interfere with endometrial innate immunity and expression of steroidogenic 49 hormone receptors at the end of the puerperium. Therefore, the objectives with this 50 study were: a) to evaluate the effect of PR and postpartum clinical mastitis on 51 reproductive efficiency, service period and interval between birth and first artificial 52 insemination (AI) of crossbred dairy cows (experiment 1); b) to evaluate the effect 53 of these diseases on innate endometrial immunity and steroidogenic hormone 54 receptors (experiment 2). For exp. 1, 232 cows were evaluated and divided into the 55 healthy, PR and clinical mastitis groups. Cows were inseminated as estrus was 56 observed or submitted to the IATF protocol. The assessments of the occurrence of 57 diseases took place after delivery until the end of the voluntary waiting period and 58 the diagnosis of pregnancy around 32 days after AI. For exp. 2, 30 cows were used, 59 divided into three groups (n = 10) equal to experiment 1. Between 40-50 days 60 postpartum clinical examination, gynecological examination and endometrial 61 sample collection by cytobrush for endometrial cytology and gene expression by 62 IL1, IL6, CCL5, ESR1 and PGR qRT-PCR technique. In exp. 1, the conception 63 rate in the first AI and the percentage of pregnant cows at 150 days were lower in 64 sick animals (PR, mastitis) and the days of the birth-first AI interval and service 65 period were higher in sick animals. In exp. 2, 26.7% (8/30) of the animals had 66 subclinical endometritis. IL1 showed higher expression in sick groups, IL6 67 showed less expression in RP group and CCL5 had no difference between groups. 68 There was no difference in the gene expression of ESR1 and PGR steroid receptors 69 between sick and healthy cows. It was concluded that cows with placenta retention 70 and clinical mastitis in the postpartum period present worse reproductive indexes, 71 alteration in the endometrial gene expression of the cytokines IL1 and IL6, 72 however, they do not present changes in the expression of steroidogenic hormone 73 receptors at the end of the puerperium.

Factores que intervienen en los mecanismos de apertura cervical en perras con piometra / Factors involved in opening cervical mechanisms in bitches with pyometra / Fatores envolvidos nos mecanismos de abertura cervical em cadelas com piometra

En Brasil, hay una gran población de perros con riesgo de desarrollar un panorama de la piometra ya que sólo un pequeño porcentaje de perros sanos se presenta generalmente a ovariohisterectomía electiva. La patogénesis de la piometra canina no se entiende completamente y puede presentar una forma abierta o cerrada, caracterizado por la presencia o ausencia de secreción vaginal, respectivamente. El relajamiento cervical es un factor importante para el diagnóstico, para establecer el prognóstico y para elegir el mejor tratamiento. La inducción de la relajación cervical, independiente del ciclo estral, también podría facilitar los examenes citológicos e histopatológicos del endométrio para el diagnóstico de procesos inflamatorios y neoplásicos. Hay numerosos factores que deben tenerse en cuenta al estudiar los mecanismos de apertura y cierre de cuello de útero, como la concentración sérica de estrógeno y progesterona, los receptores de hormonas en el útero, el porcentaje de colágeno en el cuello del útero, infiltrado inflamatorio, prostaglandinas, interleucinas y óxidos de nitrógeno. El propósito de esta revisión de la literatura es dilucidar los factores que intervienen en los mecanismos de la abertura del cuello uterino en las hembras que sufren de piometra.(AU)

In Brazil, there is a large population of female dogs at risk of developing pyometra, since only a small percentage of healthy bitches is usually submitted to elective ovariohysterectomy. Pathogenesis of canine pyometra is not fully understood and can be presented in open or closed forms characterized by the presence or absence of vaginal discharge, respectively. The cervical opening is an important factor for diagnosis, prognosis and for choosing an effective treatment. The cervical opening, regardless of the estrous cycle, would also facilitate cytological and histopathological tests for endometrial inflammatory and neoplastic processes diagnosis. Numerous factors must be considered when studying the mechanisms of cervical opening and closing, such as the serum concentration of estrogen and progesterone hormone, receptors in the uterus, the percentage of collagen in the cervix, inflammatory infiltrate, prostaglandins, interleukins and nitric oxide. The aim of this review is to elucidate the factors involved in the mechanisms of cervical opening in bitches with pyometra.(AU)

No Brasil existe uma grande população de cadelas com risco de desenvolver um quadro de piometra, pois apenas uma pequena porcentagem de cadelas sadias é normalmente submetida à ovariosalpingohisterectomia eletiva. A patogênese da piometra canina não é completamente esclarecida e pode se apresentar de forma aberta ou fechada, caracterizada pela presença ou ausência de secreção vaginal, respectivamente. O relaxamento cervical é um fator importante para o diagnostico, para o estabelecimento de um prognóstico e para a escolha de um tratamento eficiente. O relaxamento cervical, independente do ciclo estral, poderia também facilitar a realização de exames citológicos e histopatológicos do endométrio para diagnóstico de processos inflamatórios e neoplásicos. Inúmeros fatores devem ser considerados quando se estuda os mecanismos de abertura e fechamento cervical, tais como: a concentração sérica de estrógeno e progesterona, os receptores hormonais no útero, a porcentagem de colágeno na cérvix, infiltrado inflamatório, prostaglandinas, interleucinas e óxidos nítricos. O objetivo desta revisão bibliográfica é elucidar possíveis fatores envolvidos nos mecanismos de abertura cervical em cadelas portadoras de piometra.(AU)

Expressão endometrial de receptores de estradiol e progesterona em éguas cíclicas, gestantes e acíclicas tratadas com hormônios esteroidais / Endometrial expression of estradiol and progesterone receptors in cyclic, pregnant and non-cyclic steroid treated mares

A utilização de éguas receptoras cíclicas e acíclicas nos programas de transferência de embriões (TE) é uma prática consagrada na reprodução equina. Uma alternativa vantajosa para criadores e veterinários é o uso das receptoras acíclicas, uma vez que viabiliza a antecipação da estação de monta e reduz o manejo com as receptoras. Para se estabelecer a gestação nestas receptoras, estrógenos e progestinas são administrados, de forma a simular o ambiente hormonal uterino de éguas que se tornam naturalmente gestantes. A administração de estrógenos previamente às progestinas é baseada nos estudos em éguas cíclicas, que mostram que o estradiol, presente no período de estro, estimula a expressão dos seus próprios receptores e dos receptores para progesterona no endométrio. A progesterona, por sua vez, provoca mudanças uterinas que possibilitam o estabelecimento e manutenção da gestação e leva a redução dos receptores de estradiol e de seus próprios receptores durante o diestro e início da gestação. No entanto, pouco se sabe sobre os receptores endometriais de estradiol e progesterona em éguas acíclicas, principalmente utilizadas em programas de TE. O presente trabalho revisa o padrão de expressão gênica dos receptores endometriais de estradiol e progesterona em éguas durante o ciclo estral, início da gestação e submetidas a tratamentos hormonais com estrógenos e progestinas exógenas. (AU)

The use of cyclic and non cyclic recipient mares in embryo transfer (ET) programs is a well established practice in equine reproduction. An advantageous alternative for horse owners and veterinarians is the use of non-cyclic recipients, since it makes possible the anticipation of the breeding season and reduces handling with the recipients. In order to establish pregnancy in the non-cyclic recipients, estrogens and progestins are administered to allow the induction of similar endometrial changes to those which occur in naturally pregnant mares. Administration of estradiol prior to progestins is based on cyclic mares researches, where estradiol produced during estrous stimulates the expression of estrogen and progesterone receptors in the endometrium. On the other hand, progesterone causes uterine changes that allow pregnancy establishment and maintenance. In addition, progesterone leads to reduction of estrogen and progesterone receptors during diestrous in cyclic mares. However, little is known about estrogen and progesterone endometrial receptors in non-cyclic mares, especially used in ET programs. The current study revises the gene expression pattern of estradiol and progesterone endometrial receptors in mares during estrous cycle, early pregnancy and submitted to estrogens and progestins hormonal treatments. (AU)

Expressão endometrial de receptores de estradiol e progesterona em éguas cíclicas, gestantes e acíclicas tratadas com hormônios esteroidais / Endometrial expression of estradiol and progesterone receptors in cyclic, pregnant and non-cyclic steroid treated mares

A utilização de éguas receptoras cíclicas e acíclicas nos programas de transferência de embriões (TE) é uma prática consagrada na reprodução equina. Uma alternativa vantajosa para criadores e veterinários é o uso das receptoras acíclicas, uma vez que viabiliza a antecipação da estação de monta e reduz o manejo com as receptoras. Para se estabelecer a gestação nestas receptoras, estrógenos e progestinas são administrados, de forma a simular o ambiente hormonal uterino de éguas que se tornam naturalmente gestantes. A administração de estrógenos previamente às progestinas é baseada nos estudos em éguas cíclicas, que mostram que o estradiol, presente no período de estro, estimula a expressão dos seus próprios receptores e dos receptores para progesterona no endométrio. A progesterona, por sua vez, provoca mudanças uterinas que possibilitam o estabelecimento e manutenção da gestação e leva a redução dos receptores de estradiol e de seus próprios receptores durante o diestro e início da gestação. No entanto, pouco se sabe sobre os receptores endometriais de estradiol e progesterona em éguas acíclicas, principalmente utilizadas em programas de TE. O presente trabalho revisa o padrão de expressão gênica dos receptores endometriais de estradiol e progesterona em éguas durante o ciclo estral, início da gestação e submetidas a tratamentos hormonais com estrógenos e progestinas exógenas.

The use of cyclic and non cyclic recipient mares in embryo transfer (ET) programs is a well established practice in equine reproduction. An advantageous alternative for horse owners and veterinarians is the use of non-cyclic recipients, since it makes possible the anticipation of the breeding season and reduces handling with the recipients. In order to establish pregnancy in the non-cyclic recipients, estrogens and progestins are administered to allow the induction of similar endometrial changes to those which occur in naturally pregnant mares. Administration of estradiol prior to progestins is based on cyclic mares’ researches, where estradiol produced during estrous stimulates the expression of estrogen and progesterone receptors in the endometrium. On the other hand, progesterone causes uterine changes that allow pregnancy establishment and maintenance. In addition, progesterone leads to reduction of estrogen and progesterone receptors during diestrous in cyclic mares. However, little is known about estrogen and progesterone endometrial receptors in non-cyclic mares, especially used in ET programs. The current study revises the gene expression pattern of estradiol and progesterone endometrial receptors in mares during estrous cycle, early pregnancy and submitted to estrogens and progestins hormonal treatments.

Imunidade inata uterina em vacas após o parto / Uterine innate immunity in postpartum cows

A imunidade inata é o principal mecanismo de defesa responsável pelo controle da contaminaçãouterina em vacas recém-paridas. O reconhecimento inicial de determinados componentes estruturais demicrorganismos invasores do ambiente uterino é realizado por receptores específicos, localizados nas célulasendometriais. Estudos sobre a atuação desses receptores são importantes para elucidar os mecanismos queregulam a resposta endometrial contra os patógenos, a qual pode evoluir para a proteção imunológica ou para oestabelecimento de infecções uterinas nos casos de falhas na ativação da resposta inflamatória. O objetivo destarevisão é relatar as principais informações disponíveis sobre imunidade inata uterina em vacas após o parto.(AU)

Innate immunity is the main defense mechanism responsible for controlling uterine contamination inpostpartum cows. The initial recognition of certain structural components of microorganisms that invade theuterine environment is mediated by specific receptors located in endometrial cells. Studies about the role ofthese receptors are important for elucidating the mechanisms that regulate endometrial response againstpathogens, which can progress to immune protection or establishment of uterine infections in cases of failures inthe activation of the inflammatory response. The objective of this review is to describe the most relevantknowledge about uterine innate immunity in postpartum cows.(AU)

Imunidade inata uterina em vacas após o parto / Uterine innate immunity in postpartum cows

A imunidade inata é o principal mecanismo de defesa responsável pelo controle da contaminaçãouterina em vacas recém-paridas. O reconhecimento inicial de determinados componentes estruturais demicrorganismos invasores do ambiente uterino é realizado por receptores específicos, localizados nas célulasendometriais. Estudos sobre a atuação desses receptores são importantes para elucidar os mecanismos queregulam a resposta endometrial contra os patógenos, a qual pode evoluir para a proteção imunológica ou para oestabelecimento de infecções uterinas nos casos de falhas na ativação da resposta inflamatória. O objetivo destarevisão é relatar as principais informações disponíveis sobre imunidade inata uterina em vacas após o parto.

Innate immunity is the main defense mechanism responsible for controlling uterine contamination inpostpartum cows. The initial recognition of certain structural components of microorganisms that invade theuterine environment is mediated by specific receptors located in endometrial cells. Studies about the role ofthese receptors are important for elucidating the mechanisms that regulate endometrial response againstpathogens, which can progress to immune protection or establishment of uterine infections in cases of failures inthe activation of the inflammatory response. The objective of this review is to describe the most relevantknowledge about uterine innate immunity in postpartum cows.

The role of proestrus on fertility and postovulatory uterine function in the cow

Modern protocols to synchronize ovulations for timed artificial insemination and timed embryo transfer that include manipulations in the proestrus period (i.e., between luteolysis and estrus) affect fertility in cattle. Specifically, stimulating pre-ovulatory follicle growth and exposure to estrogens after CL regression increase the proportion of cows pregnant and decrease late embryo mortality. Such effects may be due to both pre-ovulatory actions of estrogens and post-ovulatory actions of progesterone, as concentrations of the later hormone may be changed in response to manipulations conducted during proestrus. In the first portion of this paper we describe strategies used recently to manipulate the proestrus period in protocols for synchronization of ovulation, and to present evidence of their effects on fertility. Manipulations of timing and prominence of sex steroids during the proestrus and early diestrus that affect fertility may act on targets such as the endometrium. This tissue expresses receptors for both estrogens and progesterone and these hormones change endometrial function to support conceptus growth and pregnancy maintenance. However, specific cellular and molecular mechanisms through which fertility is affected via manipulations of the proestrus are poorly understood. In the second portion of this paper we describe a well-defined animal model to study changes in endometrial function induced by manipulations conducted during the proestrus. Such manipulations induced endometrial changes on sex steroid receptors expression, cell proliferation, oxidative metabolism and eicosanoid synthesis in the uterus, but not on glucose transport to uterine lumen . In summary, evidence is accumulating to support a positive role of increasing duration and estrogen availability during the proestrus on fertility to synchronization protocols. Such positive effects may be through changes in endometrial function to stimulate conceptus growth and survival.